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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Ultrasound-guided transvaginal follicular aspiration to provide a source of bovine oocytes for gene microinjection

Gibbons, John R. 16 December 2009 (has links)
Three experiments were conducted to determine an efficient method of obtaining oocytes from cows via ultrasound-guided transvaginal follicular aspiration. Transvaginal oocyte recovery combined with in vitro maturation, fertilization, and culture (IVMlIVFIIVC) can produce pre-implantation stage bovine embryos and also supply a source of ova for gene microinjection. In Experiment 1, once- (IX) vs. twice-weekly (2X) oocyte recovery frequencies were compared. No differences in recovery per session were observed (IX = 6.8 vs. 2X = 6.3 oocytes/session; p>O.1 0). However, the 2X group generated more oocytes on a weekly basis (IX = 6.8 vs. 2X = 12.6 oocytes/week; p<O.OS). In Experiment 2, in vitro embryo production was compared among the groups aspirated once-weekly (1 X), twice-weekly (2X), and twice-weekly after receiving I5 mg FSH (2XF). No differences existed among the aspiration groups in the proportion of blastocysts produced following IVMIIVFIIVC (IX = 23.10/0, 2X = 26.1%, 2XF = 18.0%, % viable). However, the 2X group generated more oocytes and embryos throughout the experiment (2X = 83/318, 2XF = 38/211, IX = 58/2SI viable blastocyst/total oocytes; p<O.O5) than the other groups. A higher proportion of 2X or 2XF generated embryos were of excellent quality (2X = 60.2%, 2XF = 60.5%) compared to the IX group (37.9%) In Experiment 3, in vitro embryo development rates were compared among oocytes from follicular aspiration (TVFA) and oocytes derived from slaughterhouse ovaries (SHD). Oocytes (65%) from both sources were subjected to pronuclear-microinjection of foreign DNA while the remainder served as non-injected controls. Only control oocytes differed, with TVF A-derived oocytes developing to blastocyst more successfully than SHD oocytes (40.8% vs. 30.0olo~ p<0.05). Microinjected embryo development was similar between the groups (TVFA 15.9%, vs. SHD = 12.8% viable blastocyst/total;) with the TVFA oocytes holding a slight but non-significant numerical advantage (p>O.IO) In these experiments, twice-weekly follicular aspiration without exogenous FSH, was the best scheme of oocyte recovery. This program provided a source of consistent, high quality oocytes that responded favorable to the IVMIIVFIIVC system and microinjection. / Master of Science

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